Capillary Electrophoresis Analysis of Organic Amines and Amino Acids in Saline and Acidic Samples Using the Mars Organic Analyzer

The Mars Organic Analyzer (MOA) has enabled the sensitive detection of amino acid and amine biomarkers in laboratory standards and in a variety of field sample tests. However, the MOA is challenged when samples are extremely acidic and saline or contain polyvalent cations. Here, we have optimized the MOA analysis, sample labeling, and sample dilution buffers to handle such challenging samples more robustly. Higher ionic strength buffer systems with pKa values near pH 9 were developed to provide better buffering capacity and salt tolerance. The addition of ethylaminediaminetetraacetic acid (EDTA) ameliorates the negative effects of multivalent cations. The optimized protocol utilizes a 75mM borate buffer (pH 9.5) for Pacific Blue labeling of amines and amino acids. After labeling, 50mM (final concentration) EDTA is added to samples containing divalent cations to ameliorate their effects. This optimized protocol was used to successfully analyze amino acids in a saturated brine sample from Saline Valley, California, and a subcritical water extract of a highly acidic sample from the Río Tinto, Spain. This work expands the analytical capabilities of the MOA and increases its sensitivity and robustness for samples from extraterrestrial environments that may exhibit pH and salt extremes as well as metal ions

Ancient Microbes from Halite Fluid Inclusions: Optimized Surface Sterilization and DNA Extraction

Fluid inclusions in evaporite minerals (halite, gypsum, etc.) potentially preserve genetic records of microbial diversity and changing environmental conditions of Earth's hydrosphere for nearly one billion years. Here we describe a robust protocol for surface sterilization and retrieval of DNA from fluid inclusions in halite that, unlike previously published methods, guarantees removal of potentially contaminating surface-bound DNA. The protocol involves microscopic visualization of cell structures, deliberate surface contamination followed by surface sterilization with acid and bleach washes, and DNA extraction using Amicon centrifugal filters. Methods were verified on halite crystals of four different ages from Saline Valley, California (modern, 36 ka, 64 ka, and 150 ka), with retrieval of algal and archaeal DNA, and characterization of the algal community using ITS1 sequences. The protocol we developed opens up new avenues for study of ancient microbial ecosystems in fluid inclusions, understanding microbial evolution across geological time, and investigating the antiquity of life on earth and other parts of the solar system

The Hominin Sites and Paleolakes Drilling Project:High-Resolution Paleoclimate Records from the East African Rift System and Their Implications for Understanding the Environmental Context of Hominin Evolution

The possibility of a causal relationship between Earth history processes and hominin evolution in Africa has been the subject of intensive paleoanthropological research for the last 25 years. One fundamental question is: can any geohistorical processes, in particular, climatic ones, be characterized with sufficient precision to enable temporal correlation with events in hominin evolution and provide support for a possible causal mechanism for evolutionary changes? Previous attempts to link paleoclimate and hominin evolution have centered on evidence from the outcrops where the hominin fossils are found, as understanding whether and how hominin populations responded to habitat change must be examined at the local basinal scale. However, these outcrop records typically provide incomplete, low-resolution climate and environmental histories, and surface weathering often precludes the application of highly sensitive, state-of-the-art paleoenvironmental methods. Continuous and well-preserved deep-sea drill core records have provided an alternative approach to reconstructing the context of hominin evolution, but have been collected at great distances from hominin sites and typically integrate information over vast spatial scales. The goal of the Hominin Sites and Paleolakes Drilling Project (HSPDP) is to analyze climate and other Earth system dynamics using detailed paleoenvironmental data acquired through scientific drilling of lacustrine depocenters at or near six key paleoanthropological sites in Kenya and Ethiopia. This review provides an overview of a unique collaboration of paleoanthropologists and earth scientists who have joined together to explicitly explore key hypotheses linking environmental history and mammalian (including hominin) evolution and potentially develop new testable hypotheses. With a focus on continuous, high-resolution proxies at timescales relevant to both biological and cultural evolution, the HSPDP aims to dramatically expand our understanding of the environmental history of eastern Africa during a significant portion of the Late Neogene and Quaternary, and to generate useful models of long-term environmental dynamics in the regionpublishersversionPeer reviewe

Finishing the euchromatic sequence of the human genome

The sequence of the human genome encodes the genetic instructions for human physiology, as well as rich information about human evolution. In 2001, the International Human Genome Sequencing Consortium reported a draft sequence of the euchromatic portion of the human genome. Since then, the international collaboration has worked to convert this draft into a genome sequence with high accuracy and nearly complete coverage. Here, we report the result of this finishing process. The current genome sequence (Build 35) contains 2.85 billion nucleotides interrupted by only 341 gaps. It covers ∼99% of the euchromatic genome and is accurate to an error rate of ∼1 event per 100,000 bases. Many of the remaining euchromatic gaps are associated with segmental duplications and will require focused work with new methods. The near-complete sequence, the first for a vertebrate, greatly improves the precision of biological analyses of the human genome including studies of gene number, birth and death. Notably, the human enome seems to encode only 20,000-25,000 protein-coding genes. The genome sequence reported here should serve as a firm foundation for biomedical research in the decades ahead

Anomalously High Cretaceous Paleobrine Temperatures: Hothouse, Hydrothermal or Solar Heating?

Elevated surface paleobrine temperatures (average 85.6 °C) are reported here from Cretaceous marine halites in the Maha Sarakham Formation, Khorat Plateau, Thailand. Fluid inclusions in primary subaqueous “chevron” and “cumulate” halites associated with potash salts contain daughter crystals of sylvite (KCl) and carnallite (MgCl2·KCl·6H2O). Petrographic textures demonstrate that these fluid inclusions were trapped from the warm brines in which the halite crystallized. Later cooling produced supersaturated conditions leading to the precipitation of sylvite and carnallite daughter crystals within fluid inclusions. Dissolution temperatures of daughter crystals in fluid inclusions from the same halite bed vary over a large range (57.9 °C to 117.2 °C), suggesting that halite grew at different temperatures within and at the bottom of the water column. Consistency of daughter crystal dissolution temperatures within fluid inclusion bands and the absence of vapor bubbles at room temperature demonstrate that fluid inclusions have not stretched or leaked. Daughter crystal dissolution temperatures are reproducible to within 0.1 °C to 10.2 °C (average of 1.8 °C), and thus faithfully document paleobrine conditions. Microcrystalline hematite incorporated within halite crystals also indicate high paleobrine temperatures. We conclude that halite crystallized from warm brines rich in K-Mg-Na-Cl; sylvite and carnallite daughter crystals were nucleated during cooling of the warm brines sometime after deposition. Hothouse, hydrothermal, and solar-heating hypotheses are compared to explain the anomalously high surface paleobrine temperatures. Solar radiation stored in shallow density stratified brines is the most plausible explanation for the observed paleobrine temperatures and the progressively higher temperatures downward through the paleobrine column. The solar-heating hypothesis may also explain high paleobrine temperatures documented from fluid inclusions in other ancient halites

Starvation-Survival in Haloarchaea

Recent studies claiming to revive ancient microorganisms trapped in fluid inclusions in halite have warranted an investigation of long-term microbial persistence. While starvation-survival is widely reported for bacteria, it is less well known for halophilic archaea—microorganisms likely to be trapped in ancient salt crystals. To better understand microbial survival in fluid inclusions in ancient evaporites, laboratory experiments were designed to simulate growth of halophilic archaea under media-rich conditions, complete nutrient deprivation, and a controlled substrate condition (glycerol-rich) and record their responses. Haloarchaea used for this work included Hbt. salinarum and isolate DV582A-1 (genus Haloterrigena) sub-cultured from 34 kyear Death Valley salt. Hbt. salinarum and DV582A-1 reacted to nutrient limitation with morphological and population changes. Starved populations increased and most cells converted from rods to small cocci within 56 days of nutrient deprivation. The exact timing of starvation adaptations and the physical transformations differed between species, populations of the same species, and cells of the same population. This is the first study to report the timing of starvation strategies for Hbt. salinarum and DV582A-1. The morphological states in these experiments may allow differentiation between cells trapped with adequate nutrients (represented here by early stages in nutrient-rich media) from cells trapped without nutrients (represented here by experimental starvation) in ancient salt. The hypothesis that glycerol, leaked from Dunaliella, provides nutrients for the survival of haloarchaea trapped in fluid inclusions in ancient halite, is also tested. Hbt. salinarum and DV582A-1 were exposed to a mixture of lysed and intact Dunaliella for 56 days. The ability of these organisms to utilize glycerol from Dunaliella cells was assessed by documenting population growth, cell length, and cell morphology. Hbt. salinarum and DV582A-1 experienced size reductions and shape transitions from rods to cocci. In the short-term, these trends more closely resembled the response of these organisms to starvation conditions than to nutrient-rich media. Results from this experiment reproduced the physical state of cells (small cocci) in ancient halite where prokaryotes co-exist with single-celled algae. We conclude that glycerol is not the limiting factor in the survival of haloarchaea for thousands of years in fluid inclusions in halite

Climate reconstructions from the Tibet Plateau (PKDB286426)

Fluid inclusions of protogenous halite, which were collected from two boreholes in the Charhan Salt Lake in the north part of the Qinghai-Xizang Plateau, werea nalyzed for their hydrogen and oxygen isotopes and for their Na, Mg etc. ions.On these grounds, the evolution of lake environment in this region during the last 50 000 years are discussed in this paper. The emphasis is to discuss the time range of extremely arid and cold climate at the last Glacial stage and the geological event of playa associated with such a climate.The guanidine hydrochloride method was used for measurement of hydrogen and oxygen stable isotopes. The measurement of Na, Mg etc. ions were achieved by determination of crystallization temperature of hydrohalite under microscope and then by calculation of chemical compositions of inclusion fluid using a thermodynamic model.The results obtained show that protogenous halite in the Charhan Lake area was formed in three different environment conditions: (1) In fluid inclusions of halite formed in the early period (50 000-30 000 a B. P. ), dD averages -14.9 per mil, d(18)O averages 8.37 per mil, and Mg(2+)ranges from 0.42 to 1.59 mol/L. Their plotting points fall on the right top part of the evaporation line of the present Charhan Lake area, indicating that the Lake water at that time had a higher concentration of brine, and the climate was hot and dry. (2) In fluid inclusions of halite formed in the middle period (30 000-15 000 a B. P.), SD average -66.0 per mil, d(18)O averages 1.00 pr mil, and Mg(2+) 1 mol/L. Their plotting points fall on the left low part of the evaporation line, indicating that the lake water at that time had a concentration of brine lower than that in the early period, and the environment was cold and dry. (3) In fluid inclusions of halite formed in the late period (15 000-present), dD averages 30.8 per mil, d(18)O averages 5.85 per mil, and Mg(2+) M 1 mol/L. Their plotting fall on the evaporation line, indicating that the climate environment at that time was warm and dry, almost the same as the present.The temperature variation of the last 50 000 years in the Charhan Lake area was calculated using the conversion equation proposed by Lorious et al. The time range of the Great ice age of the Last Glacial Stage is about 21 000-15 000 a B.P., which basically coincides with the time of a worldwide low sea level. The temperature in that period was below 0°C and 6-7°C lower than now. Because of lower temperatures, water supply to the lake area decreased rapidly and the concentration of lake water increased sharply. Therefore the Mg(2+) concentration in inclusion fluid reaches or closes to 2mol/L and the Mg/Na ratio varies within a very wide range. These show that the Charhan Lake at that time entered its playa stage. The Charhan Salt Lake is a typical one in the north part of the Qinghai-Xizang Plateau. It can be supposed that the extremely arid and cold climate of the Great Ice Age made most lakes in the north part of the Qinghai-Xizang Plateau enter their playa stage. This event is of importance for formation of salt resources